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대장균 배양 중 phe W$^+$-pheS-$^{-ts}$ System에 의한 재조합 trp$^+$ 플라스미드의 안정적 유지
강충민,최장원,이세영 한국미생물 · 생명공학회 1990 한국미생물·생명공학회지 Vol.18 No.1
재조합 pBR322-trp$^+$ 플라스미드의 숙주내 안정적 유지를 목적으로 tRNA phe 의 구조유전자인 pheW$^+$ 유전자를 pBR322-trp$^+$의 플라스미드에 도입시키고, 숙주세포로는 트립토판 생산을 위한 정상숙주 LC901의 phenylalanyl tRNA synthetase 온도감수성 변이체인 LC901-pheS-ts를 구성하여 이 온도감수성 숙주의 제한온도 (restrictive temperature)에서 재조합 trp$^+$ 플라스미드의 안정적 유지와 trp$^+$ 유전자가 미치는 효과를 조사하였다. To improve the stability of recombinant pBR322-trip$^+$ plasmid (pLTW24) in E. coli culture, a positive selection system was devised. A DNA fragment containing pheW$^+$ gene (a structural gene for tRNA$^{phe}$ was isolated and inserted into the pBR322-trip$^+$ plasmid(pLTP24). A temperature sensitive host strain. LC901-pheS$^{-ts}$, was constructed for this plasmid by transducing pheS$^{-ts}$ allele (phenylalanyl-tRNA synthetase) to E. coli LC901 using P1kc bacteriophage. The LC901-pheS$^{-ts}$ cells were unable to grow at a restrictive temperature when they had lost the pBR322 :: pheW$^+$ (pLTP24) plasmid. The effects of pheW$^+$ gene on the plasmid stability and the expression level of trip$^+$ gene in LC901-pheS$^{-ts}$ strain were investigated. The proportion of Trip$^+$ colonies among LC901-pheS$^{-ts}$ strain carrying plasmid pLTP24 was 99%, whereas that of LC901 strain carrying plasmid pLTW24 was 7% at the end of 20 generations. After 100 generations of growth, the strain LC901-pheS$^{-ts}$ carrying plasmid pLTP24 showed little loss of plasmids. While the majority of plasmid pLTW24 in LC901 strain were lost in the same period. The activities of tryptophan synthetase (T. Sase) and anthranilate synthetase (A. Sase) in LC901 strain carrying pLTW24 were about 1.2 times and 1.8 times respectively of those in LC901-pheS$^{-ts}$ strain carrying pLTP24.
서울시 미세입자 (PM2.5)의 호흡기질환 사망과의 연관성 연구
강충민,박성균,선우영,강병욱,이학성 한국대기환경학회 2006 한국대기환경학회지 Vol.22 No.5
Numerous epidemiological studies have shown stronger associations between PM2.5 and both mortality and morbidity than PM10. The association of PM2.5 with respiratory mortality was examined in Seoul, during the period of 1996~2002. Because PM2.5 data were available for only 10% of this time period, a prediction regression model was developed to estimate PM2.5 concentration. Death count due to respiratory-related diseases (total respiratory mortality; ICD-10, J00-J98) and death counts (cause-specific mortality) due to pneumonia (ICD-10, J12-J18), COPD (ICD-10, J40-J44) and asthma (ICD-10, J45-J46) were considered in this study. Averaged daily mortality was 5.6 for total respiratory mortality and 1.1 to 1.6 for cause-specific mortality. Generalized additive Poisson models controlling for confounders were used to evaluate the acute effects of particle exposures on total respiratory mortality and cause-specific mortality. An IQR increase in 5-day moving average of PM2.5 (22.6 μg/m3) was associated with an 8.2% (95% CI: 4.5 to 12.1%) increase in total respiratory mortality. The association of PM2.5 was stronger for the elderly (≥65 years old, 10.1%, 95% CI: 5.8 to 14.5%) and for males (8.9%, 95% CI: 2.1 to 11.3%). A 10-μg/m3 increase in 5-day moving average of PM2.5 was strongly associated with total respiratory mortality in winter (9.5%, 95% CI: 6.6 to 12.4%), followed by spring (3.1%, 95% CI: -1.2 to 7.5%), which was a different pattern with the finding in North American cities. However, our results are generally consistent with those observed in recent epidemiological studies, and suggest that PM2.5 has a stronger effect on respiratory mortality in Seoul.
Kang, Chung Min,Choi, Jang Won,Lee, Se Yong 한국산업미생물학회 1990 한국미생물·생명공학회지 Vol.18 No.1
재조합 pBR322-trp^+ 플라스미드의 숙주내 안정적 유지를 목적으로 tRNA^phe의 구조유전자인 pheW^+ 유전자를 pBR322-trp^+ 플라스미드에 도입시키고, 숙주 세포로는 트립토판 생산을 위한 정상숙주 LC901의 phenylalanyl tRNA synthetase 온도감수성 변이체인 LC901-pheS^-ts를 구성하여 이 온도감수성 숙주의 제한온도(restrictive temperature)에서 재조합 trp^+ 플라스미드의안정적 유지와 trp^+ 유전자와 발현 정도에 pheW^+ 유전자가 미치는 효과를 조사하였다. 20세대 후, LC901-pheS^-ts 균주내의 pLTP24 플라스미드는 99%가 보존된 반면 LC901 균주내의 pLTW24 플라스미드는 단지 7%만이 남아 있었다. LC901의 pLTW24 플라스미드의 경우 40세대 후에는 플라스미드가 전혀 남아 있지 않았으나, LC901-pheS^-ts의 pLTP24 플라스미드는 100세대 후에도 거의 안정하게 유지되었다. LC901 숙주내의 pLTW24 플라스미드에 의해 발현되는 tryptophan synthetase와 anthranilate synthetase의 활성도는 LC901-pheS^-ts내의 pLTP24 플라스미드 것보다 각각 1.2와 1.8배로 높게 나타났다. To improve the stability of recombinant pBR322-trp^+ plasmid(pLTW24)in E. coli culture, a positive selection system was devised. A DNA fragment containing pheW^+ gene(a structural gene for tRNA^phe) was isolated and inserted into the pBR322-trp^+ plasmid (pLTP24). A temperature sensitive host strain, LC0901-pheS^-ts, was constructed for this plasmid by transducing pheS^-ts allele(phenylalanyl-tRNA synthetase) to E. coli LC901 using Plkc bacteriophage. The LC901-pheS^-ts cells were unable to grow at a restrictive temperature when they had lost the pBR322::pheW^+-trp^+(pLTP24) plasmid. The effects of pheW^+gene on the plasmid stability and the expression level of trp^+ gene in KC901-pheS^-ts strain were investigated. The proportion of Trp^+ colonies among LC901-pheS^-ts strain carrying plasmid pLTP24 was 99%, whereas that of LC901 strain carrying plasmid pLTW24 was 7% at the end of 20 generations. After 100 generations of growth, the strain LC901-pheS^-ts carrying plasmid pLTP24 showed little loss of plamids, while the majority of plasmid pLTW24 in LC901 strain were lost in the same period. The activities of tryptophan synthetase(T. Sase) and anthranilate synthetase(A. Sase) in LC901 strain carrying pLTW24 were about 1.2 times and 1.8 times respectively of those in LC901-pheS^-ts strain carrying pLTP24.